High spatial resolution inorganic scintillator detector for high-energy X-ray beam at small field irradiation.

PURPOSE: Small field dosimetry for radiotherapy is one of the major challenges due to the size of most dosimeters, for example, sufficient spatial resolution, accurate dose distribution and energy dependency of the detector. In this context, the purpose of this research is to develop a small size scintillating detector targeting small field dosimetry and compare its performance with other commercial detectors. METHOD: An inorganic scintillator detector (ISD) of about 200 µm outer diameter was developed and tested through different small field dosimetric characterizations under high-energy photons (6 and 15 MV) delivered by an Elekta Linear Accelerator (LINAC). Percentage depth dose (PDD) and beam profile measurements were compared using dosimeters from PTW namely, microdiamond and PinPoint three-dimensional (PP3D) detector. A background fiber method has been considered to quantitate and eliminate the minimal Cerenkov effect from the total optical signal magnitude. Measurements were performed inside a water phantom under IAEA Technical Reports Series recommendations (IAEA TRS 381 and TRS 483). RESULTS: Small fields ranging from 3 × 3 cm2 , down to 0.5 × 0.5 cm2 were sequentially measured using the ISD and commercial dosimeters, and a good agreement was obtained among all measurements. The result also shows that, scintillating detector has good repeatability and reproducibility of the output signal with maximum deviation of 0.26% and 0.5% respectively. The Full Width Half Maximum (FWHM) was measured 0.55 cm for the smallest available square size field of 0.5 × 0.5 cm2 , where the discrepancy of 0.05 cm is due to the scattering effects inside the water and convolution effect between field and detector geometries. Percentage depth dose factor dependence variation with water depth exhibits nearly the same behavior for all tested detectors. The ISD allows to perform dose measurements at a very high accuracy from low (50 cGy/min) to high dose rates (800 cGy/min) and was found to be independent of dose rate variation. The detection system also showed an excellent linearity with dose; hence, calibration was easily achieved. CONCLUSIONS: The developed detector can be used to accurately measure the delivered dose at small fields during the treatment of small volume tumors. The author's measurement shows that despite using a nonwater-equivalent detector, the detector can be a powerful candidate for beam characterization and quality assurance in, for example, radiosurgery, Intensity-Modulated Radiotherapy (IMRT), and brachytherapy. Our detector can provide real-time dose measurement and good spatial resolution with immediate readout, simplicity, flexibility, and robustness.

Assessment of radioactivity in Italian baby food.

A set of measurements have been conducted to determine the activity-level of natural and artificial radionuclides in some baby foods commercialized in Italy. The measurements have been carried out using liquid scintillation, gamma, alpha and mass spectrometry. The activity concentrations ranged from 0.005 to 0.238, from 0.0082 to 1.65, from 0.0003 to 0.015 and from <13.6 to 233.3 Bq kg-1 for 210Po, 238U, 232Th and 40K respectively, whereas they are below the detection limit for 137Cs and 226Ra. The annual effective dose due to intake of 210Po, 238U, 232Th and 40K ranged from 280 and 800 µSv y-1 for infant 1 year old. These values lie well within the typical worldwide range of dose due to the ingestion of all natural radiation reported by UNSCEAR and they are below the internationally recommended level. This indicates that the baby food available in Italy would not pose any significant radiological impact to infant.

A normal phase high performance liquid chromatography method for the separation of hydroxy and non-hydroxy neutral lipid classes compatible with ultraviolet and in-line liquid scintillation detection of radioisotopes.

In this paper, we report a method for the separation of hydroxy fatty acid and non-hydroxy fatty acid containing neutral lipid classes via normal phase HPLC with UV detection on a PVA-Sil column. The hexane/isopropanol/methanol/water based method separates all the neutral lipids in 21 min, and subsequently flushes through the polar lipids by 27 min such that prefractionation of neutral and polar lipids are not required, and the column is re-equilibrated for the next run in 15 min, for a total run time of 45 min per sample. The separation was demonstrated at both 1.0 mL/min and 1.5 mL/min for added applicability for fraction collection or inline analysis. Separation of various hydroxy fatty acid containing lipids was demonstrated from three different plant species Ricinus communis, Physaria fendleri, and engineered Arabidopsis thaliana. Additionally, we have combined this method with an in-line liquid scintillation counter for the separation and quantification of 14C labeled lipids obtained from in vivo metabolic flux experiments conducted in the developing seeds of Arabidopsis thaliana.

Determination of the gross activity of uranium, plutonium, americium and strontium in environmental samples using solid-state scintillation.

Rapid determination of selected gross alpha and beta emitters in environmental matrices by solid-state scintillation technique is discussed. This method is based on sample treatment using microwave reactor and direct measurement of digested products using powder scintillator and alkaline solution as a substitute for traditional liquid scintillation cocktail. The selected group of radionuclides was chosen with respect to their use in nuclear industry, high radiotoxicity, and the possibility of potential misuse. The work aimed at verifying the connection of microwave decomposition using alkaline solution with solid-state scintillation using a powder scintillator YAP:Ce together with an alkaline medium.

Determination of (210)Pb and (210)Po in water using the extractive scintillation cocktail Polex™.

Method validation was performed to achieve the accreditation for our determination method of (210)Pb and (210)Po in water. A Pb(NO3)2 carrier is added to the sample and lead is precipitated with Na2SxH2O. (210)Po is co-precipitated and the extractive scintillation cocktail Polex(™) is used to determine (210)Po and (210)Pb. Uranium is also extracted by Polex(™). It can be removed by washing the precipitate with 1% HNO3. The ingrowth of (210)Pb from (222)Rn during transportation time must be calculated. It has to be subtracted from the original (210)Pb in the sample and taken into account for the calculation of the lower limit of detection.

Determination of selected natural radionuclide concentrations in southwestern Caspian groundwater using liquid scintillation counting.

Twenty-seven spring water samples from southwestern shore of the Caspian Sea were analysed for their contents of gross alpha and beta, ²²²Rn, ²²6Ra and total uranium using a liquid scintillation counter (LSC). Other methods such as the Environmental Protection Agency (EPA) method 900 for gross alpha/beta measurement, radon emanation for ²²6Ra determination and laser fluorimetry for total uranium content were applied to make a comparison with the LSC technique. The levels of measured ²²²Rn and ²²6Ra range from 0.5 to 54 Bq l⁻¹ and from 14 to 297 mBq l⁻¹, respectively. The levels of gross alpha and beta are from 16 mBq l⁻¹ to 1.0 Bq l⁻¹ and from 22 to 630 mBq l⁻¹, respectively. The total uranium contents are from 3 to 66 mBq l⁻¹. It has been shown that the simple liquid scintillation counting technique could be applied conveniently for such studies where analysis of numerous samples is concerned.

Direct LSC method for measurements of biofuels in fuel.

Direct liquid scintillation counting (LSC) for quantification of biofuels content in fuels was implemented and validated on three liquid fossil fuel matrices-ethanol, gasoline and diesel. Fatty acid methyl esters (FAMEs), hydrogenated vegetable oils (HVO) and bio-ethanol were used as biofuels. The method is applicable in the range up to 100% for all tested combinations of bio components. The sensitivity and precision of the method are suitable for determination of bio component content in the blends which is appearing on the global market. The method does not require special equipment for sample preparation.

Liquid scintillation based quantitative measurement of dual radioisotopes (3H and 45Ca) in biological samples for bone remodeling studies.

Acute and prolonged bone complications associated with radiation and chemotherapy in cancer survivors underscore the importance of establishing a laboratory-based complementary dual-isotope tool to evaluate short- as well as long-term bone remodeling in an in vivo model. To address this need, a liquid scintillation dual-label method was investigated using different scintillation cocktails for quantitative measurement of (3)H-tetracycline ((3)H-TC) and (45)Ca as markers of bone turnover in mice. Individual samples were prepared over a wide range of known (45)Ca/(3)H activity ratios. Results showed that (45)Ca/(3)H activity ratios determined experimentally by the dual-label method were comparable to the known activity ratios (percentage difference ∼2%), but large variations were found in samples with (45)Ca/(3)H activity ratios in range of 2-10 (percentage difference ∼20-30%). Urine and fecal samples from mice administered with both (3)H-TC and (45)Ca were analyzed with the dual-label method. Positive correlations between (3)H and (45)Ca in urine (R=0.93) and feces (R=0.83) indicate that (3)H-TC and (45)Ca can be interchangeably used to monitor longitudinal in vivo skeletal remodeling.

Development and evaluation of a multiple-plate fraction collector for sample processing: application to radioprofiling in drug metabolism studies.

Microplate scintillation counters are utilized routinely in drug metabolism laboratories for the off-line radioanalysis of fractions collected during HPLC radioprofiling. In this process, the current fraction collection technology is limited by the number of plates that can be used per injection as well as the potential for sample loss due to dripping or spraying as the fraction collector head moves from well to well or between plates. More importantly, sample throughput is limited in the conventional process, since the collection plates must be manually exchanged after each injection. The Collect PAL, an innovative multiple-plate fraction collector, was developed to address these deficiencies and improve overall sample throughput. It employs a zero-loss design and has sub-ambient temperature control. Operation of the system is completely controlled with software and up to 24 (96- or 384-well) fraction collection plates can be loaded in a completely automated run. The system may also be configured for collection into various-sized tubes or vials. At flow rates of 0.5 or 1.0 mL/min and at collection times of 10 or 15s, the system precisely delivered 83-µL fractions (within 4.1% CV) and 250-µL fractions (within 1.4% CV), respectively, of three different mobile phases into 12 mm × 32 mm vials. Similarly, at a flow rate of 1 mL/min and 10s collection times, the system precisely dispensed mobile phase containing a [(14)C]-radiolabeled compound across an entire 96-well plate (% CV was within 5.3%). Triplicate analyses of metabolism test samples containing [(14)C]buspirone and its metabolites, derived from three different matrices (plasma, urine and bile), indicated that the Collect PAL produced radioprofiles that were reproducible and comparable to the current technology; the % CV for 9 selected peaks in the radioprofiles generated with the Collect PAL were within 9.3%. Radioprofiles generated by collecting into 96- and 384-well plates were qualitatively comparable; however, the peak resolution was greater in the profiles that were collected in 384-well plates due to the collection of a larger number of fractions per minute. In conclusion, this new and innovative fraction collector generated radioprofile results that were comparable to current technology and should provide a major improvement in capacity and throughput for radioprofiling studies.

Characterization of PI3K class IA isoforms with regulatory subunit p55alpha using a scintillation proximity assay.

Differential activation of the phosphoinositide 3-kinase (PI3K)/AKT pathway has been linked to cancer. Activation occurs through gene amplification and activating mutations. High-frequency mutations in the gene encoding the p110alpha catalytic subunit of PI3K (PIK3CA) have been observed in a variety of tumors including colon, brain, breast, ovarian, and gastric. Inhibition of PI3K kinase activity may provide a specific way to treat multiple types of human cancer. A scintillation proximity assay (SPA) was developed to detect phosphatidylinositol 3-kinase catalytic activity. Using this assay format, steady-state kinetic parameters were compared for the PI3K class IA enzymes p110alpha, p110beta, and p110delta, each coexpressed with the regulatory subunit p85alpha or splice variant p55alpha. Inhibition by the natural product wortmannin and LY294002 was detected with potencies consistent with alternate assay formats. Other biochemical assay formats have been described for phosphoinositide 3-kinases but each has its unique limitations. The simple, inexpensive, sensitive high-throughput nature of the SPA format has advanced our knowledge of isoform-specific enzymology and will facilitate the discovery of novel PI3K inhibitors.

SAPHIRE (scintillator avalanche photoconductor with high resolution emitter readout) for low dose x-ray imaging: spatial resolution.

An indirect flat panel imager (FPI) with programmable avalanche gain and field emitter array (FEA) readout is being investigated for low-dose and high resolution x-ray imaging. It is made by optically coupling a structured x-ray scintillator, e.g., thallium (Tl) doped cesium iodide (CsI), to an amorphous selenium (a-Se) avalanche photoconductor called high-gain avalanche rushing amorphous photoconductor (HARP). The charge image created by the scintillator/HARP (SHARP) combination is read out by the electron beams emitted from the FEA. The proposed detector is called scintillator avalanche photoconductor with high resolution emitter readout (SAPHIRE). The programmable avalanche gain of HARP can improve the low dose performance of indirect FPI while the FEA can be made with pixel sizes down to 50 microm. Because of the avalanche gain, a high resolution type of CsI (Tl), which has not been widely used in indirect FPI due to its lower light output, can be used to improve the high spatial frequency performance. The purpose of the present article is to investigate the factors affecting the spatial resolution of SAPHIRE. Since the resolution performance of the SHARP combination has been well studied, the focus of the present work is on the inherent resolution of the FEA readout method. The lateral spread of the electron beam emitted from a 50 microm x 50 microm pixel FEA was investigated with two different electron-optical designs: mesh-electrode-only and electrostatic focusing. Our results showed that electrostatic focusing can limit the lateral spread of electron beams to within the pixel size of down to 50 microm. Since electrostatic focusing is essentially independent of signal intensity, it will provide excellent spatial uniformity.

Screening for phosphodiesterase inhibitory activity of Thai medicinal plants.

INTRODUCTION: Phosphodiesterases (PDEs) are a group of enzymes that have powerful effects on cellular signaling because they regulate the second messenger, cAMP or cGMP. PDE inhibitors have been used for treatment of many indications such as cardiovascular diseases, chronic obstructive pulmonary diseases, erectile dysfunction and pulmonary hypertension. THE AIM OF THE STUDY: The aim of the study was to search for sources of PDE inhibitors from Thai biodiversity. MATERIALS AND METHODS: Some Thai medicinal plants used as aphrodisiac and neurotonic agents together with plants from Leguminosae collected from the North of Thailand were screened for PDE inhibitory activity using a radioassay. RESULTS: Seven from nineteen aphrodisiac and neurotonic plants as well as three from twelve Leguminosae plants showed potent PDEs inhibitory activity. The concentrations that could inhibit 50% PDE activity (IC(50)) of the active extracts were determined in comparison to the standard inhibitor, 3-isobutyl-1-methylxanthine (IBMX). Betula alnoides, Hiptage benghalensis, Leea indica and Senna surrattensis showed IC(50) values in the range of microgram per milliliter while IBMX standard showed an IC(50) value of 0.68+/-0.13 microg/ml. CONCLUSION: Thai biodiversity was the great sources of PDE inhibitors.

Methods to identify and characterize inhibitors of bacterial RNA polymerase.

RNA polymerase is essential to the viability of bacteria in all phases of growth and development and is a proven chemotherapeutic target as the cellular target of the rifamycin class of antibiotics. However, despite the characterization of multiple different classes of natural products that selectively target bacterial RNA polymerase, and the identification of a limited number of synthetic compound inhibitors, only agents of the rifamycin class have been developed and approved for human clinical use as antibiotics. Herein we describe a scintillation proximity assay (SPA) for identifying and characterizing inhibitors of bacterial RNA polymerases and that is applicable to de novo drug discovery programs through application of automated high-throughput screening methods. In addition, we describe gel electrophoresis-based methods that are applicable to the detailed characterization of inhibitors of transcriptional initiation or elongation by bacterial RNA polymerases.

Identification of non-peptidic neuromedin U receptor modulators by a robust homogeneous screening assay.

AIM: To develop a homogeneous binding assay for high-throughput screening (HTS) of hit compounds at human neuromedin U receptor (hNMU-R) 1 and to identify non-peptidic small molecule hNMU-R modulators through functional assessments and structure-activity relationship (SAR) analyses. METHODS: Membrane preparations of Chinese hamster ovary cells (CHO-K1) stably expressing hNMU-R1, [125I]hNMU-25, and wheat germ agglutinin-coupled microbeads were used to develop an HTS assay based on scintillation proximity assay (SPA) technology. This method was applied to a large-scale screening campaign against a diverse library of 36,000 synthetic compounds or natural products and subsequent confirmation studies. CHO-K1 cells stably expressing full-length hNMU-R1 or hNMU-R2 and a calcium-sensitive dye were employed to functionally measure intracellular calcium mobilization upon ligand stimulation. Preliminary SAR was determined based on limited structural modifications. RESULTS: The Ki value (0.7 nmol/L) of hNMU-25 (the natural ligand) at hNMU-R1 measured by the SPA method was consistent with that reported in the literature, and the Z'factor for this HTS assay was 0.81. A total of 100 hits, showing more than 30% competitive inhibition on [125I]hNMU-25 binding to hNMU-R1, were identified initially, 3 of which were confirmed thereafter to have reasonable hNMU-R1-binding affinities and similar chemical structures. Based on their common molecular skeleton, 203 analogs were synthesized and tested. Among the 16 analogs that retained variable hNMU-R1- binding abilities, 2 elicited calcium influx in both hNMU-R1 and hNMU-R2-expressing cells, but none displayed antagonist activity. CONCLUSION: The homogeneous hNMU-R1 binding assay is an efficient and robust tool for screening potential hNMU-R modulators. Two non-selective hNMU-R agonists discovered are of low molecular weight nature with novel chemical structures. The preliminary SAR investigation suggests that both the triphenyl and guanidinol groups are crucial to the bioactivities observed.

Monte Carlo investigations of megavoltage cone-beam CT using thick, segmented scintillating detectors for soft tissue visualization.

Megavoltage cone-beam computed tomography (MV CBCT) is a highly promising technique for providing volumetric patient position information in the radiation treatment room. Such information has the potential to greatly assist in registering the patient to the planned treatment position, helping to ensure accurate delivery of the high energy therapy beam to the tumor volume while sparing the surrounding normal tissues. Presently, CBCT systems using conventional MV active matrix flat-panel imagers (AMFPIs), which are commonly used in portal imaging, require a relatively large amount of dose to create images that are clinically useful. This is due to the fact that the phosphor screen detector employed in conventional MV AMFPIs utilizes only approximately 2% of the incident radiation (for a 6 MV x-ray spectrum). Fortunately, thick segmented scintillating detectors can overcome this limitation, and the first prototype imager has demonstrated highly promising performance for projection imaging at low doses. It is therefore of definite interest to examine the potential performance of such thick, segmented scintillating detectors for MV CBCT. In this study, Monte Carlo simulations of radiation energy deposition were used to examine reconstructed images of cylindrical CT contrast phantoms, embedded with tissue-equivalent objects. The phantoms were scanned at 6 MV using segmented detectors having various design parameters (i.e., detector thickness as well as scintillator and septal wall materials). Due to constraints imposed by the nature of this study, the size of the phantoms was limited to approximately 6 cm. For such phantoms, the simulation results suggest that a 40 mm thick, segmented CsI detector with low density septal walls can delineate electron density differences of approximately 2.3% and 1.3% at doses of 1.54 and 3.08 cGy, respectively. In addition, it was found that segmented detectors with greater thickness, higher density scintillator material, or lower density septal walls exhibit higher contrast-to-noise performance. Finally, the performance of various segmented detectors obtained at a relatively low dose (1.54 cGy) was compared with that of a phosphor screen similar to that employed in conventional MV AMFPIs. This comparison indicates that for a phosphor screen to achieve the same contrast-to-noise performance as the segmented detectors approximately 18 to 59 times more dose is required, depending on the configuration of the segmented detectors.

Development of a sensitive radiorespiration method for detecting microbial activity at subzero temperatures.

We have developed a simple and sensitive method to detect microbial respiration at subzero temperatures. Microbial activity was detected by measuring (14)CO(2) evolved during the microbial-mediated mineralization of [1-(14)C] acetic acid or [2-(14)C] glucose in microcosm assays using modified (14)CO(2) traps. Various (14)CO(2) traps, designed to withstand freezing at subzero temperatures, were tested for their quench characteristics during liquid scintillation spectrometry and their ability to trap (14)CO(2). Solutions consisting of 1 M KOH supplemented with 20% or 30% v/v ethylene glycol did not freeze at temperatures above -20 degrees C and had a minor quenching effect on liquid scintillation spectrometry. Addition of ethylene glycol did have an effect on the efficiency of (14)CO(2) trapping, as the cumulative recovery of (14)CO(2) was reduced by 14% and 32% in the 1 M KOH+20% ethylene glycol and 1 M KOH+30% ethylene glycol solutions, respectively. Using the modified (14)CO(2) traps, microbial activity in representative Canadian high Arctic environmental samples was detected at temperatures as low as -15 degrees C. This simple method allows for sensitive, specific, and reliable detection of microbial activity occurring at subzero temperatures and is readily adaptable for studies in other cryoenvironments.

A cell-free scintillation proximity assay for studies on lysosome-to-phagosome targeting.

Phagocytes, such as macrophages, neutrophils, and dendritic cells, play important roles in the innate immune system through their ability to engulf, kill, and digest invading microbes. In cooperation with the humoral adaptive immune system, coating of substrates with immunoglobulin G (IgG) antibodies enhances several aspects of phagocytosis, including the recognition of substrates by cell surface IgG (Fcgamma) receptors, particle internalization, generation of microbicidal oxygen species, and targeting of lysosomes to phagosomes. We describe a cell-free scintillation proximity assay developed to study the mechanisms of lysosome targeting to phagosomes and the regulation of this process by IgG. The approach involves the use of isolated phagosomes containing scintillant latex beads and lysosomes labeled with a tritiated marker. Scintillation results only when lysosomes and phagosomes come into immediate contact and requires supplementation of reactions with adenosine triphosphate and cytosol; addition of cytosol from IgG-conditioned cells enhances this signal. The method is useful for investigating the biochemistry and regulation of the early tethering and docking steps of lysosome and phagosome interactions.

A simple strategy for mitigating the effect of data variability on the identification of active chemotypes from high-throughput screening data.

Among the several goals of a high-throughput screening campaign is the identification of as many active chemotypes as possible for further evaluation. Often, however, the number of concentration response curves (e.g., IC(50)s or K(i)s) that can be collected following a primary screen is limited by practical constraints such as protein supply, screening workload, and so forth. One possible approach to this dilemma is to cluster the hits from the primary screen and sample only a few compounds from each cluster. This introduces the question as to how many compounds must be selected from a cluster to ensure that an active compound is identified, if it exists at all. This article seeks to address this question using a Monte Carlo simulation in which the dependence of the success of sampling is directly linked to screening data variability. Furthermore, the authors demonstrate that the use of replicated compounds in the screening collection can easily assess this variability and provide a priori guidance to the screener and chemist as to the extent of sampling required to maximize chemotype identification during the triage process. The individual steps of the Monte Carlo simulation provide insight into the correspondence between the percentage inhibition and eventual IC(50) curves.

Performance characteristics of sequential separation and quantification of lead-210 and polonium-210 by ion exchange chromatography and nuclear spectrometric measurements.

A selective separation and quantitative determination procedure for 210Pb and 210Po in various environmental matrices from different sources such as IAEA-326 soil, phosphate rocks (PR), and phosphogypsum (PG) was developed. The tested samples were digested sequentially using concentrated mineral acids (HF, HNO3) by a programmable high-pressure microwave digestion system. The sample solution was loaded onto a preconditioned ion exchange column (Sr-resin) for chromatographic separation. Polonium-210 was eluted by 6 M HNO3 then spontaneously deposited onto polished silver discs to be measured using low-background alpha spectrometry. Lead-210 was sequentially eluted using 6 M HCl solution, precipitated as lead oxalate, dissolved in HNO3 solution, and mixed with scintillation cocktail to be measured by liquid scintillation counting (LSC). Performance of the developed procedure was tested using a reference soil (IAEA-326), with recommended isotope values, that was used as a quality control to assess separation and quantification efficiency (recovery %). The minimum detectable activities of 210Pb and 210Po were found to be 24 and 0.28 Bq kg(-1) for the measurements using LSC and alpha spectrometry, respectively. The recoveries (%) of 210Pb and 210Po were found to be 80 and 60%, respectively. To test the validity of the proposed LSC method, a comparative study was performed by measuring 210Pb activity concentration in test samples by nondestructive gamma-ray spectrometry.

A quantitative high-throughput trapping assay as a measurement of potential for bioactivation.

Idiosyncratic adverse drug reactions (ADRs) are one of the most common causes of pharmaceutical withdrawals and labeling changes. Most ADRs are caused by drugs that form reactive species that can bind covalently to macromolecules such as proteins. The current methodology for the measurement of covalent binding relies on the use of radiolabeled material that requires an investment in time and resources not typically expended until later in the discovery process. Efforts are also made to identify reactive intermediates by the use of chemical trapping agents, such as reduced glutathione and cyanide, to form stable adducts that are characterized by liquid chromatography-tandem mass spectrometry and/or nuclear magnetic resonance spectroscopy. Here, we describe a high-throughput assay for the measurement of reactive intermediate formation. The method involves incubation of cold compound with liver microsomes in the presence of [14C]potassium cyanide. Hard electrophilic species would react with the trapping agent, resulting in the formation of a radiolabeled conjugate. Unreacted trapping agent is removed using solid-phase extraction, and the amount of radiolabeled conjugate present is determined by liquid scintillation counting. This newly developed screen has proved to be specific, sensitive, robust, and a powerful tool for assessing bioactivation potential.